Research Article Natural Medicinal Materials 43

GC/MS analysis and antimicrobial activity of essential

oils of Telekia speciosa (Schreb.) Baumg.
E RMINA C ILOVI Ć KOZAREVI Ć1,* , B ROZA Š ARI Ć -K UNDALI Ć1 , M ERIMA I BIŠEVI Ć1 , E MIR H OROZI Ć2 ,
J ASMINA G LAMO ČLIJA3 , M ARINA S OKOVI Ć3 , J ELENA A RSENIJEVI Ć4 , AND Z ORAN M AKSIMOVI Ć4
1 Facultyof Pharmacy, University of Tuzla, Urfeta Vejzagića 8, 75000 Tuzla, Bosnia and Hercegovina
2 Facultyof Technology, University of Tuzla, Urfeta Vejzagića 8, 75000 Tuzla, Bosnia and Hercegovina
3 Department of Plant Physiology, Institute for Biological Research "Siniša Stanković"-National Institute of Republic of Serbia, University of Belgrade, Bulevar

Despota Stefana 142, 11000 Belgrade, Serbia

2 Department of Pharmacognosy, Faculty of Pharmacy, University of Belgrade, Vojvode Stepe 450, 11221 Belgrade, Serbia
* Corresponding author: ermina.cilovic@untz.ba

Published: December 25, 2021

Received: April 19, 2021
Accepted: July 8, 2021
Published on-line: November 30, 2021

Telekia speciosa (Schreb.) Baumg., Asteraceae, is widespread in Eastern and Central Europe and the
Balkan Peninsula. Previous phytochemical investigations have revealed T. speciosa as a rich source of
sesquiterpene lactone – isoalantolactone, especially in its underground parts. The aim of the present study
was to analyze the essential oils from aerial and underground parts of T. speciosa and investigate their
antimicrobial activity. Chemical composition of essential oils was determined by GC-FID/MS method
leading to the identification of 67 compounds in total, with 15.77 % oxygenated monoterpenes, 7.77 %
sesquiterpene hydrocarbons, 49.14 % oxygenated sesquiterpenes, and 12.37 % other compounds from
aerial parts, and 3.80 % oxygenated monoterpenes, 3.13 % sesquiterpene hydrocarbons, 90.33 % oxygenated
sesquiterpenes from underground parts essential oil. The main components from aerial parts were
(E)-nerolidol (11.54 %) and caryophyllene oxide (10.54 %), while isoalantolactone was the predominant
component from essential oil underground parts (83.41 %). The minimum inhibitory concentration
(MIC), minimum bactericidal/fungicidal concentration of the essential oils were evaluated against six
strains of bacteria and two strains of fungus using in vitro microdilution method. Both oils presented
antimicrobial properties against pathogens Staphylococcus aureus, Bacilus cereus, Pseudomonas aeruginosa,
Escherichia coli, and Candida albicans. Inhibition of growth of tested microorganisms by T. speciosa
underground parts essential oil was achieved with MICs ranging from 1.0 to 11.0 mg mL-1 , while MICs of
aerial parts essential oil varied from 4.0 to 30.0 mg mL-1 . The obtained results contribute to the knowledge
of antimicrobial properties of T. speciosa, which support traditional uses underground parts of the plant.

Key words: Telekia speciosa; Asteraceae; essential oil composition; antimicrobial activity; isoalantolactone

http://dx.doi.org/10.5937/leksir2141043C

1. INTRODUCTION to genus Inula, including Inula helenium L., a well-known medic-

Telekia speciosa (Schreb.) Baumg. (Asteraceae) is a perennial
herbaceous plant widespread in Eastern and Central Europe
and the Balkan Peninsula. It is up to 200 cm high and it has
alternating, wide, whole leaves and large heterogeneous yellow
flowers with a diameter of 5-8 cm which can be individual or
in cluster inflorescence. It inhabits wet and shady positions in
mountain woodlands (Chalchat et al., 2004; Domac, 1994), and
somewhere it is planted as a decorative plant. It is closely related
inal plant (Stojakowska et al., 2011). The underground parts
of T. speciosa are traditionally used as a remedy for bronchial
asthma in Balkan countries (Serbia, Bosnia and Herzegovina).
For example, smoke after burning underground parts is inhaled
for the cure of asthma (Redzić, 2007).
The underground parts of T. speciosa contain essential oil, bitter
compounds and inulin (Marković et al., 2010). Phytochemical in-
vestigations have revealed T. speciosa as a rich source of sesquiter-
 Research Article
pene lactone – isoalantolactone, especially in its underground
parts which amount is equivalent to I. helenium underground
parts (Radulović et al., 2010; Stojakowska et al., 2011; 2015a).
The aerial parts extracts have been found to contain fatty acids,
namely palmitic, linoleic, oleic, and caproic acids, and sterols
(Deliorman et al., 2002; Orhan and Sener, 2003). Aerial parts of
T. speciosa accumulated miscellaneous sesquiterpene lactones,
mainly of guaiane, pseudoguaiane, xanthane, and eudesmane
type (Stojakowska et al., 2015b).
Isoalantolactone, a bioactive compound present in T. speciosa
underground parts and aerial parts essential oils, has been
found to have various pharmacological activities including anti-
inflammatory, antimicrobial, and anticancer properties, with no
significant toxicity. Alantolactone and isoalantolactone have
been extensively investigated on several cancer cell lines, such
as colon, melanoma, ovary, prostate, lung, and leukemia (Rasul
et al., 2013). A previous study also revealed that extracts from
both leaves and flowers of T. speciosa showed high antiprolif-
erative activity against the cancer cell lines tested (Yuan et al.,
2018). In addition, several in vitro and in vivo studies evaluated
antimicrobial properties of isoalantolactone against methycillin-
resistant Staphylococcus aureus (MRSA) and α-toxin, a product of
most S. aureus microorganisms and essential for the pathogen-
esis of pneumonia (Qiu et al., 2011; Zhou et al., 2020). Unlike
T. speciosa, for which, as far as we know, there are no available
data on antimicrobial activity, essential oil of I. helenium un-
derground parts was active against several Gram-positive and
Gram-negative bacteria and Candida strains (Deriu et al., 2008).
Increasing bacterial resistance to antibiotics, antimicrobials, and
antifungal agents is a growing concern facing the medical, phar-
maceutical, sanitation, and food industries (Krist et al., 2015). An
alternative to reduce the use of synthetic chemicals is the search
for antimicrobials from medicinal plants. In this context, plants
constituents, such as essential oils and their main components,
terpenoids, have attracted considerable interest (Swamy et al.,
2016).
The aim of the present study was to analyze the essential oils of
both, aerial and underground parts of T. speciosa and to evaluate
antimicrobial activity of essential oils.
2. MATERIALS AND METHODS
2.1. Plant material
Aerial and underground parts of T. speciosa were collected at the
location of Karaula, Olovo municipality (Bosnia and Herzegov-
ina) (N44°10’22.3", E18°38’58.6") during the flowering period in
July 2018. The plant material was identified according to Flora
Croatica (Domac, 1994) by authors and the voucher specimen
was deposited at the Department of Pharmacognosy, Faculty
of Pharmacy, the University of Tuzla. The plant material was
cleaned, cut, and air-dried.
2.2. Isolation and GC-FID/MS analyses of volatiles
The dried aerial and underground parts of T. speciosa were
chopped and subjected to hydrodistillation for 3 h using a
Clevenger-type apparatus. The obtained essential oils were
separated, dried over anhydrous sodium sulfate, and stored at
-20 °C until the analysis. The constituents of essential oils were
determined by the GC-FID/MS method, as described previously
(Cilović et al., 2019).
Natural Medicinal Materials 44
2.3. Antimicrobial activity of essential oils
The antimicrobial activity (AMA) of T. speciosa essential oils
were tested against six strains of bacteria, i.e. Gram-positive
Staphylococcus aureus (ATCC 6538 and clinical isolate) and Bacil-
lus cereus (clinical isolate), and Gram-negative Pseudomonas aerug-
inosa (ATCC 27853 and clinical isolate) and Escherichia coli (ATCC
35210), as well as against two strains of fungus Candida albicans
(ATCC 10231 and clinical isolate). The microorganisms were
obtained from the Mycology Laboratory of the Institute for Bi-
ological Research “Siniša Stanković”, University of Belgrade,
Serbia.
The antibacterial assay was done by microdilution method
(Cazella et al., 2019) utilizing 96-well microtiter plates to deter-
mine the minimal inhibitory concentration (MIC) and minimal
bactericidal /fungicidal concentration (MBC/MFC). The inocu-
lum was cultivated in a solid medium to verify the absence of
contaminations, and for validation. Nutrient media were Tryp-
tic Soy Broth (TSB) for bacteria and Sabouraud Dextrose Broth
(SDB) for fungi. The microorganism suspensions (inocula) were
adjusted with sterile saline until the concentration of 1.0 x 105
CFU mL-1 . The inoculum was prepared daily and stored at 4 °C
until its utilization. The essential oil was added to the nutrient
medium for the growth of microorganisms. Then the microor-
ganism inocula were added and the plates were incubated for
24 h at 37 °C for bacteria and 72 h at 28 °C for fungi. The lowest
concentration without visible microbial biomass growth under a
binocular magnifying glass was defined as MIC.
Determining the absence of growth of microorganisms, i.e. de-
termination MBC/MFC, was performed by serial reinoculation
of 10 µL of inoculated medium from wells where no growth of
the microorganism was recorded in 100 µL of sterile nutrient
medium and reincubation for 24 h at 37 °C for bacteria and for
72 h at 28 °C for fungi. The results were confirmed after adding
40 µL of purple p-iodonitrotetrasolium chloride (Sigma), micro-
bial growth indicator solution (0.2 mg mL-1 distilled water) to
each well and incubation for 30 minutes at 37 °C (Tsukatani
et al., 2012). Comparison of color intensity was performed with
control wells in which unhindered growth of microorganisms
was enabled, and commercial antimicrobial agents streptomycin
(Sigma), ampicillin (Panfarma, Belgrade, Serbia), and ketocona-
zole (Zorka farma, Šabac, Serbia) were used as positive controls.
Inoculated medium without added essential oil was used as the
negative control. The antimicrobial tests were carried out in
triplicate. The results were expressed in values of arithmetical
average ± standard deviation.
3. RESULTS AND DISCUSSION
3.1. Content and composition of essential oils
The aerial parts of T. speciosa contained 0.02 % (v/w) of yellow,
liquid fragrant essential oil. The identified 49 constituents ac-
counting for 85.05 % of the oil are presented in Table 1. The
essential oil of aerial parts (AEO) of T. speciosa was characterized
by the presence of a high concentration of oxygenated sesquiter-
penes (49.14 %). The major components were (E)-nerolidol (11.54
%), caryophyllene oxide (10.54 %), and (2Z,6E)-farnesol (4.52 %).
Sesquiterpene hydrocarbons constituted 7.77 % of the oil with
the dominant compound β-caryophyllene (4.90 %). Nerol (4.77
%) was the major representative of oxygenated monoterpenes,
which constituted 15.77 % of the oil. Non-terpene (other) com-
pounds presented an appreciable amount of essential oil (12.37
%) with dominant (E)-phytol (4.14 %).
Research Article Natural Medicinal Materials 45

Table 1. Chemical composition of essential oils from T. speciosa aerial and underground parts
No. Ret. time Compound RIEa AEOb UEOc
[%m/m] [%m/m]
1 12.844 linalool 1101.1 3.15 0.15
2 15.109 nerol oxide 1155.2 0.52 -
3 16.674 α-terpineol 1192.5 0.66 -
4 17.251 n-decanal 1206.2 0.84 -
5 18.236 nerol 1229.3 4.77 0.09
6 20.660 bornyl acetate 1286.0 0.47 -
7 20.852 dihidroedulan I 1290.4 2.48 -
8 22.325 silphiperfol-5-ene 1325.1 - 0.21
9 23.124 7-epi-silphiperfol-5-ene 1343.7 - 0.36
10 23.704 eugenol 1357.4 0.44 -
11 24.496 silphiperfol-6-ene 1376.4 - 0.12
12 24.682 modheph-2-ene 1380.9 - 0.14
13 24.947 α-isocomene 1387.2 - 0.12
14 25.199 β-elemene 1393.2 - 0.21
15 25.250 phenyl ethyl isobutanoate 1394.8 0.41 -
16 25.758 β-isocomene 1406.7 0.38 0.2
17 26.383 β-caryophyllene 1422.6 4.9 0.44
18 26.398 2,5-dimethoxy-p-cymene 1422.8 0.28 0.1
19 27.387 epi-β-santalene 1447.5 - 0.22
20 27.639 geranyl acetone 1454.0 0.91 -
21 27.710 α-humulene 1456.0 0.46 -
22 28.584 β-chamigrene 1477.7 - 0.16
23 28.893 thymol isobutyrate 1485.7 0.76 0.24
24 29.019 (E)-β-jonone 1488.8 1.11 -
25 29.099 β-selinene 1490.7 - 0.46
26 29.153 neryl isobutanoate 1492.1 3.07 0.77
27 29.397 α-selinene 1498.1 0.67 0.31
28 29.809 β-bisabolene 1508.6 - 0.18
29 30.009 modhephen-8-β-ol 1514.3 0.61 0.35
30 30.235 (E)-dihydro-apofarnesal 1519.3 0.39 -
31 30.479 δ-cadinene 1526.0 0.82 -
32 31.240 α-calacorene 1545.7 0.54 -
33 31.589 isocaryophyllene oxide 1554.8 1.33 -
34 31.815 epi-longipinanol 1560.6 0.57 -
35 32.053 (E)-nerolidol 1566.4 11.54 -
36 32.410 neryl (S)-2-metilbutyrate 1575.8 - 0.48
37 32.425 prenopsan-8-ol 1576.2 4.7 -
38 32.945 caryophyllene oxide 1589.7 10.54 0.98
39 33.795 humulene epoxide II 1612.4 1.71 0.14
40 34.488 murola-4,10(14)-diene-1-β-ol 1631.2 0.33 -
41 34.582 α-acorenol 1634.0 - 0.22
42 34.676 caryophylla-4(12),8(13)-dien-5-α-ol 1636.3 2.04 -
43 35.320 β-eudesmol 1653.8 2 0.66
44 35.565 selin-11-en-4-α-ol 1660.8 0.88 0.44
45 35.753 14-hydroxy-9-epi-β-caryophyllene 1665.8 - 0.2
46 35.923 14-hydroxy-(Z)- caryophyllene 1670.5 - 0.14
47 36.107 14-hydroxy-9-epi-(E)- caryophyllene 1675.6 0.89 0.53
48 36.418 epi-α-bisabolol 1683.7 1.31 -
49 36.655 eudesma-3,11-(13)-dien-12-al 1690.6 0.55 0.21
50 36.954 n-heptadecane 1698.6 0.49 -
51 37.218 δ-dodecalactone 1705.8 0.29 -
52 37.510 (2E,6Z)-farnesal 1714.6 0.66 -
53 37.678 3-methoxy-cuminil isobutyrate 1719.1 - 0.22
54 37.795 (2Z,6E)-farnesol 1722.8 4.52 -
55 37.887 naphthalen-2-ol 1725.3 - 0.49
56 38.523 (2E,6E)-farnesal 1743.4 0.47 -
57 38.867 ciclocolorenone 1753.7 0.39 -
58 39.224 (Z)-lanceol 1763.7 0.46 -
59 40.435 n-octadecane 1798.6 0.29 -
60 43.894 alantolactone 1902.7 - 2.56
61 44.280 10-isobutyryloxi thymol isobutyrate 1914.6 - 0.33
62 44.410 (5E,9E)-farnesyl acetone 1919.0 0.33 -
63 44.416 10-isobutyryloxy-8.9-dehydrothymyl isobutyrate 1919.2 0.34 -
64 45.229 isoalantolactone 1944.7 3.64 83.41
65 47.940 10-isobutyryloxy-8,9-epoxithymyl isobutyrate 2031.6 1.31 1.42
66 50.427 (E)-phytol 2114.3 4.14 -
67 60.949 n-pentacosane 2498.9 0.69 -
Total identified 85.05 97.26
Oxygenated monoterpenes 15.77 3.8
Sesquiterpene hydrocarbons 7.77 3.13
Oxygenated sesquiterpenes 49.14 90.33
Others 12.37 -
a
RI, retention indices as determined on HP-5 column using homologous series of C8 -C30 alkanes.
b
AEO, aerial parts essential oil.
c
UEO, underground parts essential oil
 Research Article
The amount of essential oil found in the underground parts
(UEO) of T. speciosa (0.31 %, v/w) was higher than in the aerial
parts. The oil was yellowish semi-solid mass, with aromatic odor.
By cooling and standing it crystallizes in the form of opaque
needle-like crystals. The identified 35 constituents accounting
for 97.26 % of the oil are presented in Table 1. This essential
oil was characterized by the presence of a high concentration of
oxygenated sesquiterpenes (90.33 %) with isoalantolactone (IAL)
being the major component (83.41 %). Alantolactone, an oxy-
genated sesquiterpene, was also presented, but in a substantially
lower amount (2.56 %). Sesquiterpene hydrocarbons constituted
3.13 %, while oxygenated monoterpenes constituted 3.80 % of
the oil, with thymol derivative 10-isobutyryloxi-8,9-epoxithymol
isobutyrate (1.42 %), found in appreciable amount.
In the previous investigations, aerial parts collected during the
flowering period of T. speciosa from Bosnia and Herzegovina
and Serbia contained slightly higher amounts of essential oil
than in the current study, i.e. 0.04 % and 0.06 % (v/w), respec-
tively. The major components of those oils were oxygenated
sesquiterpenes (E,Z)-farnesol (12.0 % in the essential oil from
Serbia), (E)-nerolidol (10.2-10.3 %), caryophyllene oxide (4.5-8.2
%), (Z,E)-farnesol (7.7 % in the essential oil from Bosnia and
Herzegovina), β-caryophyllene (5.4 %), similar to the currently
analyzed AEO (Cilović et al., 2019; Radulović et al., 2010).
It is worth noting that T. speciosa leaf essential oil, originating
from Poland, as dominant constituents also contained a far-
nesol isomer (i.e. (E,E)-farnesol, 21.2 %) and (E)-nerolidol (17.9
%), while flower essential oil was abundant with isoalantolac-
tone (23.0 %) and 10-isobutyryloxy-8,9-epoxythymol isobutyrate
(20.5 %) along with other several thymol derivatives (Wajs-
Bonikowska et al., 2012). These two compounds were present
in several times lower amounts in the currently analyzed oil,
which could be explained by the fact that it was obtained from
the complete aerial parts in which flowers represented only one
minor part.
Based on the previously and currently analyzed essential oil
from T. speciosa aerial parts it could be concluded that the charac-
teristic compounds for analyzed oil are (E)-nerolidol, caryophyl-
lene oxide, different stereoisomers [(E,Z)-, (Z,E)-, (E,E)-] of far-
nesol, β-caryophyllene, and other compounds present in lower
amounts.
In the previous investigations, underground parts collected dur-
ing the flowering period of T. speciosa from Bosnia and Herze-
govina and Poland contained almost equal or somewhat higher
amounts of essential oil than in the current study, i.e. 0.29 %
and 0.41 % (v/w), respectively. Only underground parts of T.
speciosa collected during the flowering period from Montenegro
contained higher amounts of essential oil than in the current
study, i.e. 1.7 %. The dominant compound in all those samples
was isoalantolactone (62.3 % - 95 %). Beside isoalantolactone,
other oxygenated sesquiterpene alantolactone (2.4 % in the es-
sential oil from Bosnia and Herzegovina), and thymol deriva-
tives 10-isobutyryloxi-8,9-epoxithymol isobutyrate (2.9-3.4 %), 9-
isobutyryloxythymol isobutyrate (2.1 % in the essential oil from
Poland) were also present in the previously analyzed essential
oils (Chalchat et al., 2004; Cilović et al., 2019; Wajs-Bonikowska
et al., 2012). Results about the chemical composition of T. speciosa
underground parts essential oil obtained in this study are in ac-
cordance with the published literature data. Taking into account
previous data and the currently analyzed oil underground parts
of T. speciosa may be considered as a potential raw material for
isoalantolactone isolation.
Natural Medicinal Materials 46
3.2. Antimicrobial activity of essential oils
Both investigated essential oils AEO and UEO exerted antimi-
crobial activity against all tested microorganisms (Tables 2 and
3).
As shown in Table 2, bacterio- and fungistatic effects of UEO
were more prominent then those of AEO, but lower then the
control antibiotics streptomycin and ampicillin and antifungal
ketoconazole. Inhibition of growth of tested microorganisms by
T. speciosa UEO was achieved with MICs ranging from 1.0 to 11.0
mg mL-1 , while MICs of AEO varied from 4.0 to 30.0 mg mL-1 .
Similarly, bactericide and fungicide effects were observed in the
presence of lower concentrations of UEO (MBC/MFC 4.0-15.0
mg mL-1 ) in comparison to AEO (MBC/MFC 7.0-90.0 mg mL-1 )
(Table 3). However, both essential oils were less effective than
the antimicrobial drugs that were used as positive controls.
Among tested bacteria, ATCC strains of S. aureus and E. coli were
the most susceptible strains to T. speciosa UEO (MIC 1.1, and 1.0
mg mL-1 , respectively; MBC 7.0 mg mL-1 for both microorgan-
isms), while both standard and clinical strains of P. aeruginosa
were the most (and equally) resistant in the presence of this
oil (MIC and MBC were 7.0 mg mL-1 , and 15.0 mg mL-1 , re-
spectively). There was no apparent difference in susceptibility
toward UEO between Gram-positive and Gram-negative bacte-
ria. On the other hand, AEO exhibited stronger effects on the
tested Gram-negative bacteria than on Gram-positive ones. In
addition, even though AEO showed generally lower activity
in comparison to UEO, the standard strain of P. aeruginosa (the
only one among the tested strains) was more susceptible to AEO
(MIC 4.0 mg mL-1 ) than to UEO (MIC 7.0 mg mL-1 ). The most
resistant strain to T. speciosa AEO was a clinical isolate of B. cereus
(MIC 30.0 mg mL-1 ; MBC 90.0 mg mL-1 ). Clinical isolate of C.
albicans was highly susceptible to UEO (MIC 1.0 mg mL-1 ), but
not to AEO (MIC 11.0 mg mL-1 ). Both EOs showed similar and
relatively low anticandidal activity on standard strain of this
fungus.
The analyzed essential oils did not show a large difference in
AMA against clinical and ATCC strains of microorganisms, in
contrast, to control antibiotics (streptomycin, ampicillin) and
antifungal ketoconazole which showed a lower effect on clinical
than on standard strains, except in the case of C. albicans. Clinical
isolate of this fungus is more than 10 times susceptible to UEO
compared to the standard strain.
The potential of major constituents in the currently analyzed
oils to act as antimicrobials or to contribute to the antimicro-
bial effects of essential oils was confirmed in several previous
studies. Qiu et al. (2011) investigated the activity of IAL on
S. aureus and α-toxin. α-Toxin is a product of most S. aureus
microorganisms and is essential for the pathogenesis of pneu-
monia. MIC value of IAL for S. aureus was more than 1.024 mg
mL-1 and IAL inhibited the expression of α-toxin, in S. aureus
at very low concentrations. Results were confirmed in vitro and
in vivo. Also IAL, in combination with penicillin G, exhibited
significant synergism against 21 β-lactamase-positive S. aureus
strains (including MRSA). MIC values of the penicillin G alone
and in combination with IAL against tested bacterial S. aureus
and MRSA strains were reduced three to even twenty-six times
(Zhou et al., 2020).
Nerolidol, the major compound in the currently analyzed T.
speciosa AEO, is common component found in the essential oil
of various medicinal plants. Many studies showed its antimi-
crobial activity. The mixture of (Z,E)-nerolidol exhibited potent
antimicrobial activity against S. aureus and 20 strains of MRSA
 Research Article Natural Medicinal Materials 47

Table 2. Minimum inhibitory concentrations (MIC) of Telekia speciosa aerial and underground parts essential oils and
streptomycin, ampicillin and ketoconazole controls
Microorganism AEOa UEOb Streptomycin Ampicillin Ketoconazole
[mg mL-1 ] [mg mL-1 ] [mg mL-1 ] [mg mL-1 ] [mg mL-1 ]
Staphylococcus aureus ATCC 6538 15.0 ± 2.0 1.1 ± 0.1 0.006 ± 0.001 0.012 ± 0.001 n.d.
Staphylococcus aureus clinical isolate 15.0 ± 1.0 4.0 ± 0.3 0.100 ± 0.010 0.100 ± 0.008 n.d.
Bacillus cereus clinical isolate 30.0 ± 3.0 3.0 ± 0.1 0.025 ± 0.003 0.100 ± 0.010 n.d.
Pseudomonas aeruginosa ATCC 27853 4.0 ± 0.3 7.0 ± 0.5 0.025 ± 0.002 0.050 ± 0.002 n.d.
Pseudomonas aeruginosa clinical isolate 11.0 ± 1.0 7.0 ± 0.6 0.100 ± 0.015 0.300 ± 0.009 n.d.
Escherichia coli ATCC 35210 7.0 ± 0.5 1.0 ± 0.1 0.100 ± 0.002 0.150 ± 0.008 n.d.
Candida albicans ATCC 10231 15.0 ± 1.0 11.0 ± 1.0 n.d. n.d. 0.0016 ± 0.0001
Candida albicans clinical isolate 11.0 ± 2.0 1.0 ± 0.1 n.d. n.d. 0.0031 ± 0.0001
a
AEO, aerial parts essential oil.
b
UEO, underground parts essential oil.
c
n.d., not detected.

Table 3. Minimum bactericidal/fungicidal concentrations (MBC/MFC) of Telekia speciosa aerial and underground parts
essential oils and streptomycin, ampicillin, and ketoconazole controls.
Microorganism AEOa UEOb Streptomycinc Ampicillin Ketoconazole
[mg mL-1 ] [mg mL-1 ] [mg mL-1 ] [mg mL-1 ] [mg mL-1 ]
Staphylococcus aureus ATCC 6538 30.0 ± 2.0 7.0 ± 0.5 0.012 ± 0.001 0.025 ± 0.002 n.d.
Staphylococcus aureus clinical isolate 30.0 ± 1.2 7.0 ± 0.3 0.200 ± 0.01 0.150 ± 0.009 n.d.
Bacillus cereus clinical isolate 90.0 ± 5.0 4.0 ± 0.1 0.050 ± 0.006 0.150 ± 0.01 n.d.
Pseudomonas aeruginosa ATCC 27853 7.0 ± 1.0 15.0 ± 1.0 0.050 ± 0.009 0.100 ± 0.007 n.d.
Pseudomonas aeruginosa clinical isolate 15.0 ± 1.0 15.0 ± 1.5 0.200 ± 0.01 0.500 ± 0.03 n.d.
Escherichia coli ATCC 35210 15.0 ± 2.0 7.0 ± 0.6 0.200 ± 0.008 0.200 ± 0.02 n.d.
Candida albicans ATCC 10231 30.0 ± 2.0 15.0 ± 1.0 n.d. n.d. 0.0062 ± 0.001
Candida albicans clinical isolate 15.0 ± 1.5 7.0 ± 0.5 n.d. n.d. 0.0058 ± 0.300
a
AEO, aerial parts essential oil.
b
UEO, underground parts essential oil.
c
n.d., not detected.

(Hada et al., 2003). (E)-Nerolidol also exhibited antimicrobial
activity against S. aureus with MIC values ranging from 125
to 500 µg mL-1 (Braca et al., 2008). In another study MIC val-
ues for nerolidol against Streptococcus mutans, Salmonela enterica,
and Aspergillus niger were 25.0, from 3.9 to 15.6, and 62.5 µg
mL-1 , respectively (Chan et al., 2016). For (E,E)-farnesol, ali-
cyclic sesquiterpene alcohol present in many essential oils as
well as in T. speciosa AEO, good antimicrobial and fungistatic
activity is reported (Krist et al., 2015).
Even though MIC values for caryophyllene oxide and β-
caryophyllene, to the best of knowledge, were not yet published,
the AMA of essential oil from leaves of Croton heliotropiifolius
which contained β-caryophyllene as dominant compound inhib-
ited the growth of several Gram-positive and Gram-negative
bacteria. Its antibacterial activity was characterized as weak to
moderate against the analyzed strains (de Alencar Filho et al.,
2017).
In the context of the current investigation, it is worth noting
that there is a great similarity in the chemical composition of
the essential oil of T. speciosa underground parts with previously
analyzed essential oil of Inula helenium L. root which exhibited
substantial antimicrobial activity against several bacterial and
fungal strains. The dominant principles of I. helenium root es-
sential oil were three isomeric eudesmane-type sesquiterpene
lactones: alantolactone (51.3-55.8 %), isoalantolactone (26.1-36.9
%), and diplophyllin (5.1 %). It was experimentally confirmed
that essential structural parts responsible for antimicrobial activ-
ity of I. helenium root essential oil are eudesmane core olefinic
bonds, alongside the α,β-methylene-lactone ring of sesquiter-
pene lactones. Bacterial and fungal strains used in this research
were similar to those in the current study. Clinical isolate of
B. cereus and standard strain of S. aureus were among the most
susceptible strains to analyzed I. helenium L. root essential oil
(MIC 0.017 and 0.6 mg mL-1 , respectively). The most resistant
strains were clinical isolate of P. aeruginosa and standard strain
of E. coli (MIC 14.8 mg mL-1 for both). Analyzed essential oil
showed generally better AMA against Gram-positive strains
regarding Gram-negative ones. Candida strains were among the
most susceptible with MIC values ranging from 0.009 mg mL-1
to 0.12 mg mL-1 (Deriu et al., 2008; Stojanović-Radić et al., 2012).
T. speciosa UEO obtained best results of antibacterial activity also
for the standard strain of S. aureus, clinical isolate of B. cereus,
and unlike I. helenium root essential oil, for the standard strain
of E. coli. Clinical isolate of P. aeruginosa was among the most
resistant strains to this essential oil, too. The oil showed also
good fungicidal activity against clinical isolate of C. albicans,
while standard strain was among the most resistant ones.
CONCLUSION
In the current study, Telekia speciosa (Schreb.) Baumg. was char-
acterized by the presence of a high concentration of oxygenated
sesquiterpenes with isoalantolactone as the predominant con-
stituent of essential oil underground parts, and (E)-nerolidol
 Research Article
and caryophyllene oxide as the major constituents of essential
oil aerial parts. According to this, underground parts may be a
potential raw material for isoalantolactone isolation.
T. speciosa essential oil was, to the best of our knowledge, investi-
gated for antimicrobial activity against pathogens for the first
time in this study. Essential oil of underground parts achieved
better antimicrobial activity compared to the essential oil of
aerial parts with the most susceptible standard strains of S. au-
reus and E. coli, and clinical isolate of C. albicans.
The obtained results are important from the aspect of T. speciosa
application as an antimicrobial agent. Traditionally its under-
ground parts are used in bronchial asthma therapy and have
a great potential to be used as a new therapeutic drug with
recommendation for further pharmacological and toxicological
investigations.
ACKNOWLEDGMENTS
This research is funded by the Serbian Ministry of Education,
Science and Technological Development (Contract No. 451-03-
68/2020-14/200007)
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